Science Bulletin 63 (2018) 133–142

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Science Bulletin
journal homepage: www.elsevier.com/locate/scib

Review

Antibacterial applications of graphene oxides: structure-activity

relationships, molecular initiating events and biosafety
Huizhen Zheng a, Ronglin Ma a, Meng Gao a, Xin Tian a, Yong-Qiang Li a, Lingwen Zeng b,c,⇑, Ruibin Li a,⇑
a
School for Radiological and Interdisciplinary Sciences (RAD-X), Collaborative Innovation Center of Radiation Medicine of Jiangsu Higher Education Institutions, Soochow
University, Suzhou 215123, China
b
Institute of Environmental and Food Safety, Wuhan Academy of Agricultural Science and Technology, Wuhan 430000, China
c
Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China

a r t i c l e i n f o a b s t r a c t

Article history: Bacterial infections may lead to diverse acute or chronic diseases (e.g., inflammation, sepsis and cancer).
Received 15 August 2017 New antibiotics against bacteria are rarely discovered in recent years, which necessitates the exploration
Received in revised form 16 November 2017 of new antibacterial agents. Engineered nanomaterials (ENMs) have been extensively studied for antibac-
Accepted 11 December 2017
terial use because of their long lasting killing effects in wide spectra of bacteria. Graphene oxide (GO) is
Available online 15 December 2017
one of the most widely studied ENMs and exhibit strong bactericidal effects. The physicochemical prop-
erties of GO play important roles in bacterial killing by triggering a cascade of toxic events. Many studies
Keywords:
have explored the signaling pathways of GO in bacteria. Although molecular initiating events (MIEs) of
2D Materials
Physicochemical properties
GO in bacteria dominate its killing efficiency as well as toxicity mechanisms, they have been rarely
Nanotoxicity reviewed. In this report, we discussed the structure–activity relationships (SARs) involved in GO-
Infection induced bacterial killing and the MIEs including redox reaction with biomolecules, mechanical destruc-
Microbe tion of membranes and catalysis of extracellular metabolites. Furthermore, we summarized the clinical or
commercial applications of GO-based antibacterial products and discussed their biosafety in mammal.
Finally, we reviewed the remaining challenges in GO for antibacterial applications, which may offer
new insights for the development of nano antibacterial studies.
Ó 2017 Science China Press. Published by Elsevier B.V. and Science China Press. All rights reserved.

1. Introduction stress response, destruction of bacterial membrane, and interac-

Bacterial infection has been considered as one of the greatest
threats to human health [1], because the wide uses of antibiotics
have led to the increasing spread of multidrug resistant bacteria
(a. k. a., superbugs) [2]. In addition, biofilm formation is another
major hurdle that affects the bacterial killing efficiency of tradi-
tional antibiotics [3]. Despite these crises, the discovery of new
antibiotics has significantly reduced during the past few years
[4], which necessitates the exploration of new antibacterial agents.
The development of engineered nanomaterials (ENMs) provides
opportunities to design a new generation of antibacterial agents as
an alternative to antibiotics [5]. To date, ENMs including CuO [5],
graphene oxide (GO) [6], Ag [7], ZnO [8], have been reported to
show bactericidal effects against broad spectra of Gram-positive
and Gram-negative bacteria. Compared to antibiotics, nano
antibacterial agents exhibit two advantages: (1) ENMs are able to
kill bacteria by multiple mechanisms (Fig. 1), including oxidative
⇑ Corresponding authors.
E-mail addresses: lzeng8@126.com (L. Zeng), liruibin@suda.edu.cn (R. Li).
tion with cytosolic molecules; (2) ENMs exhibit long lasting bacte-
ricidal effects in the prevention of bacterial growth on the surfaces
of solid substrates (e.g., paper, water filtration membrane, skin,
etc.) owing to their extraordinary stability [6]. Among these nano
antibacterial agents, GO has unique two dimensional (2D) honey-
combed hydrophobic plane structure and hydrophilic groups
including carboxylic (–COOH) and hydroxyl (–OH) groups on its
edge [9]. This amphipathic structure of GO nanosheets could well
facilitate their interactions with biomolecules including lipids, pro-
teins, DNA, etc., and induce bacterial death without intracellular
process. In contrast, some metal-based antibacterial nanoparticles
(e.g., Ag) could dissolve in biological media and the released metal
ions would diffuse into cytoplasm for bacterial killing. The physic-
ochemical properties of GO have been demonstrated to play an
important role in bacterial killing by triggering a cascade of toxic
events [10,11]. The downstream signals as well as bacterial killing
efficiency were significantly influenced by the molecular initiating
events (MIEs) that reflect the initial interactions between GO and
biomolecules in bacteria. Although different mechanisms including
nano-knife, oxidative stress, membrane disruption, have been

https://doi.org/10.1016/j.scib.2017.12.012
2095-9273/Ó 2017 Science China Press. Published by Elsevier B.V. and Science China Press. All rights reserved.
134 H. Zheng et al. / Science Bulletin 63 (2018) 133–142

Fig. 1. (Color online) Comparison of bacterial killing mechanisms between antibiotics and ENMs. Antibiotics could prevent bacteria growth by inhibiting the synthesis of
target biomolecules in bacteria, including cell walls, DNA, proteins, etc. In contrast, nanoparticles (e.g., AgNP, GO) could prevent bacterial growth by deconstruction of cell
wall/membrane, interaction with DNA and proteins, and disruption of electron transport as well as the redox state in bacteria.

reported in GO-induced bacterial death [10], the MIEs involved in
these toxicity pathways are rarely reviewed.
In this report, we summarized the MIEs of GO in bacteria and
discussed the key physicochemical properties of GO that con-
tribute in bacterial killing efficiencies as well as MIEs. Since GO is
found to induce significant toxicity in mammalian cells or animals
[12], substantial concerns on the biosafety of GO have been raised.
Therefore, we also discussed the biohazard effects of GO in mam-
mals from the perspective of speeding up its clinical and commer-
cial use in antibacterial products.
2. Structure-activity relationships (SARs) of GO in antibacterial
effects
and the number of corners and sharp edges [20]. Usually, GO with
larger lateral sizes can bind bacterial cells more easily and show
stronger bacterial killing [17]. When the lateral sizes of GO were
reduced to several nanometers (e.g., graphene quantum dots), their
antibacterial activities had a dramatic decrease due to the
increases of hydrophilicity and biocompatibility [21]. However,
Perreault et al. [16] found that smaller GO exhibited higher antimi-
crobial activity than the larger one. When the surface areas of GO
nanosheets decreased from 0.65 to 0.01 lm2, their bactericidal
effects increased four folds, which could be ascribed to the exces-
sive active defects on the surfaces of smaller GO (Fig. 2). These con-
tradictory results imply that size may be not the only contributor
for GO-induced bacterial death.

2.1. Size 2.2. Shape

GO exhibits both lateral and vertical structures, however, its lat-
eral diameters are tens to hundreds of times larger than the verti-
cal thicknesses [13–16]. Atomic force microscopy (AFM) and
transition electron microscopy (TEM) are popularly used to charac-
terize the primary sizes of GO [17], while the hydrodynamic sizes
of GO in solutions are estimated using dynamic light scattering
methods.
The layer numbers (thicknesses) of GO have been demonstrated
to significantly affect its antimicrobial activities [18]. Along with
the increasing of GO thicknesses from few to tens of nanomaterials,
its dispersibility in biological media displays a remarkable
decrease, resulting in GO agglomerate formation, which may affect
the interactions between GO and bacteria [15]. Therefore, mono-
layer GO has been reported to exhibit higher antibacterial activities
than multilayer GO [19].
The lateral sizes are also found to influence the antibacterial
activities of GO by altering its adsorption abilities, dispersibilities
Shapes are considered to directly impact the interactions
between GO nanosheets and bacterial cell membranes [22–25].
Although this assumption is difficult to be proved in experiments,
recent progresses on cloud computing and machine learning have
allowed theoretical simulations to predict the GO-bacteria interac-
tions. Li et al. [26] found that GO nanosheets with sharp corners
and edge protrusions can easily permeate into bacterial mem-
brane, because this unique structure makes the penetration pro-
cess undergoes a lower energy barrier [26]. Zeng’s group [27]
simulated the interaction between GO and cell membrane by
molecular dynamics, suggesting that the unique two-dimensional
(2D) structure enables robust adsorption of lipid molecules on
GO surface. Interestingly, a recent study highlighted the contribu-
tions of surface curvature of GO in bacterial killing. Two types of
graphene quantum dots (GQDs) derived from C60 and GO exhibited
different bacterial killing effects (Fig. 3a and b). C60-GQDs are more
potent (100% killing) against the spherical bacteria compared to
 H. Zheng et al. / Science Bulletin 63 (2018) 133–142 135

Fig. 2. (Color online) Antimicrobial activity of GO-coated filters for different GO sheet areas. (a) Cell viability of deposited and planktonic E. coli cells after 3 h of contact with
GO-coated filters, determined by Live/Dead fluorescent staining. (b) Number of viable E. coli cells after 3 h of contact with GO-coated filters, determined by CFU agar plate
counting. (c) Schematic image to explain the size effect of GO on antimicrobial activity. Reproduced with permission from Ref. [16]. Copyright 2015, American Chemical
Society.

rod-shaped bacteria, because the surface Gaussian curvature of
C60-GQDs matches better with spherical bacteria, which could
facilitate the association of GO with bacterial surface (Fig. 3c) [28].
2.3. Surface functionality
Compared to other carbonaceous nanomaterials like carbon
nanotube, GO is much more complex. Besides the unique two-
dimensional structure of GO, its basal plane is decorated with
epoxy and carbon radicals (C), while the edges have carboxyl
and hydroxyl groups [29,30]. These groups vary in proportion
due to different preparation methods. Therefore, reported antibac-
terial efficiencies of GO vary a lot in different laboratories [31–33].
Hu et al. [34] compared the antibacterial effects of GO and reduced
GO (rGO), and found GO nanosheets showed higher antibacterial
activity than rGO, because GO has more surface charges and func-
tionalities. Liu et al. [31] also found that GO delivered the strongest
antimicrobial activity compared with rGO, graphite (Gt) and gra-
phite oxide (GtO), which can be attributed to the improved disper-
sion ability. In contrast, Akhavan and Ghaderi [23,35] discovered
that rGO exhibited better charge transfer at nano-bio interfaces
and induced stronger bactericidal effects than pristine GO. Chong
et al. [36] demonstrated that light-induced electrons promote the
reduction of GO with additional carbon-centered free radicals to
enhance the antibacterial activities of GO. Although there are some
inconsistences in these studies, surface functional groups are con-
sidered to have great influences on the antibacterial activity of GO.
In order to discern the key functionalities involved in GO-induced
bactericidal effects, our group prepared a series of GO nanosheets
with different functional groups by controlling the reaction condi-
tions (time and temperature). We demonstrated that hydrated GO
(hGO) prepared in aqueous alkalized solution showed the highest
carbon radical densities and antibacterial effects, while rGO had
a lower antibacterial activity than pristine GO [37]. Although car-
bon radicals have been found to play an important role in the
antibacterial activity of GO derivatives by inducing membrane
lipid peroxidation, more investigations are required to obtain a
comprehensive understanding on the contribution of each func-
tionality (Fig. 4).
3. The MIEs involved in bactericidal pathways
3.1. Redox reaction with biomolecules
The growth and metabolic processes of bacteria involve many
chemical reactions, such as electron transfer, chemical moiety
modification and so on. The reactive functional groups on GO
136 H. Zheng et al. / Science Bulletin 63 (2018) 133–142

Fig. 3. (Color online) Contribution of surface curvature in GO-induced bacterial killing. Bacterial plate killing assays using C60-GQD (a) and GO-GQD (b); both GQDs are at 200
lg/mL in saline. After 3 h treatment, C60-GQD significantly reduces the viability of S. aureus (1: ATCC 25,923; 2: ATCC 29,213) but barely affects that of the other three bacteria
tested–B. subtilis, E. coli, P. aeruginosa–regardless of their Gram-property. In stark contrast, GO-GQD barely affects the survival of all five strains tested. (c) Schematic
illustration of surface curvature match between a GQD and a target bacterium surface may be correlated with the GQD’s antibacterial property. Reproduced with permission
from Ref. [28]. Copyright 2016, American Chemical Society.

surface may disrupt the redox reactions in bacteria by interact-
ing with biomolecules, such as proteins, lipids and DNA. We
have found that when GO binds or interacts with bacterial mem-
brane, carbon radicals on its surfaces could induce lipid peroxi-
dation, membrane damage and finally bacterial death. The
reactions between GO and membrane lipids may involve three
steps: (1) electron transfer from C to one of the C atom in unsat-
urated lipids; (2) formation of a lipid peroxide radicals via elec-
tron transfer from unsaturated lipids to bystander molecular
dioxygen (O2); (3) generation of a lipid peroxide (Fig. 5) [37].
Besides lipid peroxidation, Salas et al. [38] demonstrated that
electron transfer to GO is mediated by cytochromes expressed
in all Shewanella strains that take part in extracellular electron
transfer process.
3.2. Mechanical destruction
Physical interactions underlying the antimicrobial activity of
GO include membrane piercing [39], surface adhesion [40], wrap-
ping [41], and lipid extraction [42]. Mechanical injury to bacterial
membranes can be identified by morphological changes in cell
structures, leakages of cytosolic contents, internalizations of
membrane-impermeable dyes, and disruptions of transmembrane
potentials [43–45].
GO can interact with bacterial cell membrane to induce physi-
cal/mechanical injury by two ways. One is called ‘‘insertion mode”
or ‘‘penetration mode”, by which GO can spontaneously pierce into
cell membrane and cause cell membrane injury through its sharp
edges, a.k.a. nanoknives, cutters, or blades [39,46]. Liu et al. [31].
 H. Zheng et al. / Science Bulletin 63 (2018) 133–142 137

Fig. 4. (Color online) The effects of surface functionalities of GO on bacterial killing. The functionality density on GO are demonstrated to play a major role in bacterial death.
Although carbon radicals were found to induce lipid peroxidation in bacteria, the contributions of other surface functionalities are rarely studied.

Fig. 5. (Color online) Schematic image to explain the bactericidal effect of GO including membrane association and lipid peroxidation. Reproduced with permission from Ref.
[37]. Copyright 2016, American Chemical Society.

studied the antibacterial activities of GO, rGO, graphite (Gt) and
graphite oxide (GtO) toward E. coli, and found that sharp edges
can induce significant membrane stress in bacteria by acting as
cutters. To further explore the destruction details at nano-bio
interfaces, computational simulations were used for understanding
the molecular dynamics involved in GO-bacteria interaction.
Penetration of GO into bacterial membranes is largely dependent
on its corners or asperities [26]. Because GO has hydrophobic plane
and hydrophilic edges, this amphiphilic nanostructure enables its
transmembrane capability. In addition, the orientation and lateral
size were also found to impact the membrane penetration effi-
ciency of GO nanosheets and their antibacterial activities. Small
nanosheets with preferentially perpendicular orientation showed
enhanced penetration capacity [47]. Pham et al. [25] thought that
the antibacterial behavior of GO may result from the formation
of pores in bacterial cell walls, the osmotic imbalance and death.
Besides penetration effect, some researchers proposed another
interaction model that larger sheets may lie flat on the top of
membrane bilayers, induce a patch of upturned phospholipids
and finally lead to cell death [45]. The lipid injury by GO is usually
associated with its physical interaction with bacterial membranes.
Simulation studies showed that graphene nanosheets can pene-
trate into bacterial cell membrane and extract large amounts of
phospholipids on the surfaces [44]. Tu et al. [44] simulated the
138 H. Zheng et al. / Science Bulletin 63 (2018) 133–142

extraction processes at atomic levels and found that van der Waals
interaction is the main driving force at the beginning, and then
hydrophobic interaction plays a dominant role (Fig. 6). Extraction
of phospholipids by graphene nanosheets may further induce sev-
ere membrane damage, and cell death. Regrettably, most MIEs
studies on the physical/mechanical injury of GO are based on com-
putational simulations without direct experimental evidences. The
development of high resolution single atom imaging technology
may provide opportunities for in situ observation of the detailed
interaction processes between GO nanosheets and cell membranes.
3.3. Catalysis of extracellular metabolites
strongly depends on ambient pH, temperature, and H2O2 concen-
tration [49].
Although GO exhibits different MIEs, including redox reaction
with biomolecules, mechanical destruction of membranes and
catalysis of extracellular metabolites (Fig. 7), they may trigger sim-
ilar toxicity signaling pathways [50], such as reactive oxygen spe-
cies (ROS) generation, glutathione (GSH) depletion, protein and
DNA damages [31,32,51], and finally bacterial death [42,52]. The
MIEs of GO nanosheets may be influenced by their physicochemi-
cal properties [10]. Since the properties of GO nanosheets vary
with sample batches, it is necessary to simultaneously assess the
nano-SAR and MIEs for the same source of GO species.

Recently, Elimelech’s group [42] developed a GO functionalized 4. GO enabled antibacterial nanoproducts

probe to measure the interaction forces between GO and E. coli by
AFM. They observed significant repulsive effect between GO and
bacteria, implying that new MIEs independent of GO-membrane
association may be involved in the bactericidal pathways of GO.
Qu’s group [48] found that graphene quantum dots (GQDs) are able
to catalyze the decomposition of H2O2 to generate OH and further
induce bacterial death. This study demonstrated that the killing
effects of GO could be achieved by an indirect way, catalysis of
extracellular metabolites on GO surfaces, providing a new insight
on the MIEs of GO as antibacterial agents. Additionally, they dis-
covered that carboxyl-modified graphene oxide (GO-COOH) exhi-
bits intrinsic peroxidase-like activity and its catalysis capacity
Because GO allows diverse engineering designs and exhibits
unique antibacterial mechanisms, they have been extensively
studied to develop antibacterial nanoproducts for biomedical,
environmental and food engineering applications (Table 1) [53,54].
Since biomedical devices are some of the major routes for bacte-
rial infection [65], combination of GO with medical apparatus and
instruments, such as antibacterial textiles and coating catheters,
may provide effective protection against lethal infectious bacteria
[37,66,67]. Our group [37] has successfully prepared GO coating sil-
icone catheters, which could significantly prevent the growth of
drug-resistant bacteria colonies on catheter surfaces. Roy’s group

Fig. 6. (Color online) Simulated lipid extraction process by GO. (a–f) A representative trajectory of a fully restrained GO nanosheet docked at the surface of the outer
membrane (pure POPE). The simulation time is indicated in each snapshot, with the last snapshot shown in more view angles (e, f, rotated anticlockwise by angle from its
previous view to obtain the current view). Reproduced with permission from Ref. [44]. Copyright 2013, Nature Publishing Group.
 H. Zheng et al. / Science Bulletin 63 (2018) 133–142 139

Fig. 7. (Color online) MIEs of GO in bacteria. There are three different MIEs involved in bacterial killing mechanisms of GO, including redox reaction with biomolecules,
mechanical destruction of membranes and catalysis of extracellular metabolites. The downstream signaling pathways may include ROS generation and GSH depletion,
protein and DNA damages.

Table 1
GO-based antibacterial nanoproducts.

GO-based nanomaterial Antibacterial matrix Bacteria stains Applied fields Refs.

GO/rGO Polymer membranes E. coli Water purification [55]
Polymer membranes Environmental and daily-life [56]
Antibacterial agent Water purification [57]
Antibacterial paper Environmental and clinical [34]
Medical cloths Wound disinfection [58]
Ag@GO/rGO nanocomposite Hydrogels filter E. coli Water disinfection [59]
Antibacterial agent S. aureus, A. baumannii, E. faecalis Biomedical antibacterial [60]
Antifungal agent F. graminearum Crop disease prevention [61]
Peptide-GO Membrane S. aureus Biomedical antibacterial [62]
PVK@GO Membrane filter E. coli, B. subtilis Waste water treatment [53]
PVA-GO-Ag-Starch Polymer films E. coli, S. aureus Food packaging materials [63]
GO@CS@TiO2 Cling packaging film A. niger, B. subtilis Plastic wrap for food [64]

[58] designed rGO coated cloths, which can be potentially used as
bandage materials for the dressing of open wounds because of the
high antibacterial activity and hydrophobicity of rGO. Apart from
catheter and cloths, Zhang et al. [68] synthesized an Ag@GO
nanocomposite and incorporated it into bioceramic scaffolds by a
soaking method for both antibacterial and osteogenic purposes.
Biofouling has been considered as one of the major problems in
environmental engineering, because it induces enormous harm
upon environmental facilities and causes increased operation cost.
In water treatment, the porous ultrafiltration membranes are often
contaminated by bacteria, which may further form biofilm and
affect the efficiency of water filtration. To address this problem,
GO has been investigated for incorporation into ultrafiltration
membranes to prevent biofouling [56,69]. Micron-scale-scale GO
nanosheets as additives in porous ceramic membranes could also
successfully prevent the growth of bacterial colony on membrane
as well as biofilm formations [57,59].
GO-based antimicrobial products also have important utilities in
food industry as anti-corrosion additives or disinfectants [70,71].
The safety and high antibacterial efficiency are two important tar-
gets for additives to extend food shelf life. Kurantowicz et al. [72]
reported that a low concentration of GO has a significant antibacte-
rial effect against the food-borne bacterial pathogens, Listeria mono-
cytogenes and Salmonella enterica. GO additives in food packaging
materials not only show significant antibacterial and antioxidant
activities, but also provide extraordinary mechanical strength [63].
5. Biosafety of GO in mammals
In order to speed up the commercial or clinical use of GO, it is
important to evaluate the biosafety of GO in mammals. Despite
the advantages that GO present for antibacterial applications, such
as high surface area, persistent antimicrobial effects, and excellent
dispersion in aqueous environments, its biocompatibility remains
a challenge. Recently, there have been a number of publications
describing the potential toxic effects of GO in mammalian cells
and animals. GO has been found to induce ineligible hazard effects
in mammalian systems via plasma membrane disruption, lysoso-
mal dysfunction, or mitochondria damage, etc. (Fig. 8) [73]. Similar
140 H. Zheng et al. / Science Bulletin 63 (2018) 133–142
Fig. 8. (Color online) The toxic effects of GO in mammalian cells. GO may bind with cell membranes or internalized into lysosomes during their interaction with mammalian
cells. For the GO sheets associated with membrane, they may disrupt the integrity of membranes and lead to the leakage of cytosolic contents. For the GO trapped into
lysosomes, it may induce lysosome damages, NLRP3 inflammasome activations and inflammatory cytokine release. The released GO from lysosomes may interact with
mitochondria and trigger massive ROS generation.
to its antibacterial activity, the mammalian toxicities of GO were
mainly influenced by its concentration, lateral size and surface
chemistry [74].
5.1. Concentration
A considerable amount of reports have shown that GO induces
dose-dependent toxicity in cells and animals, such as cell
apoptosis, inflammatory effects, liver and kidney injury, lung fibro-
sis, etc. [75–78]. Low and middle doses (0.1–0.25 mg/kg) of GO had
negligible toxicity, while a high dose (0.4 mg/kg) of GO showed
chronic toxicity in mice [79]. Chang et al. [80] have reported a
dose-dependent ROS generation in GO-treated cells with a slight
loss of cell viability at high exposure concentrations. Therefore,
we recommend that viability tests should be performed on GO in
mammals to determine the threshold of GO doses suitable for their
safe use as antibacterial agents.
5.2. Size
The lateral sizes of GO may impact its cellular uptake and
biodistributions in animal organs [74]. Ma et al. [81] found that
large GO showed strong binding with plasma membrane and less
phagocytosis, eliciting robust interactions with Toll-like receptors
and NF-jB pathway activation, while small GO sheets were more
internalized into cytoplasm. In an intraperitoneal exposure study,
small GO (100–500 nm) preferentially accumulated in the liver,
whereas large GO (1–5 lm) was filtered out by the pulmonary cap-
illary vessels and mainly located in lungs [82]. Wang et al. [77]
demonstrated that large GO was the most pro-fibrogenic material
based on rapid kinetics of pulmonary injury. This result is consis-
tent with a previous report showing that intravenously injected
GO mainly accumulated in liver and lung, and large GO had a high
accumulation level in animal lungs [83]. Compared to mammalian
cells, bacteria cannot take GO into cytoplasm because of the lack of
endocytosis pathways. Small and large GO nanosheets often show
similar distribution in bacteria.
5.3. Surface chemistry
GO sheets possess widely varying surface chemistries that
would be affected by their sources and synthesis methods [84].
The ratios of functional groups are also different among prepara-
tions. The strong hydrophobic surfaces of GO enable the formation
of protein corona in biological systems, which could minimize the
physical interactions between GO and cell membranes and reduce
cytotoxicity [85]. The influences of surface charge have also been
highlighted in GO-induced toxicities because the electrostatic
repulsion between GO and non-phagocytes plays an important role
in particle internalization [74]. Since the biocompatibility of GO
could be improved by engineering design of its surface functional-
ities, it is urgent to discern the role of each specific functional group
in bacteria and mammalian systems. Considering the substantial
toxic effects of GO in animals after intravenous, intraperitoneal,
subcutaneous, or intramuscular injection [86], direct exposure of
GO-based antibacterial agents to human cells may induce ineligible
hazard effects.
6. Future perspective
6.1. Understanding the interactions at GO-bacteria interfaces
Although substantial achievements have been made in the stud-
ies on bactericidal mechanisms of GO, there are some challenges in
understanding the detailed information about GO-bacteria interac-
tions, e.g., how to experimentally reveal the molecular dynamics in
the interactions between GO and bacterial membrane and what is
the contribution of each physicochemical properties in GO-
induced bacterial death. Since the initial interactions at nano-bio
interfaces are the vital links between the physicochemical
properties of ENMs and their bio-effects, future studies on GO as
antibacterial agents should be more focused on the MIEs. These
studies may offer new insights to overcome the bottlenecks in
microbiology, e.g., superbugs and biofilm formation. In addition to
GO, a variety of nanosheets including MoS2, SnSe, WS2, BN, etc. have
 H. Zheng et al. / Science Bulletin 63 (2018) 133–142
been explored during the past few years [87,88]. The elements in
these materials are densely packed in an atomic-scale pattern and
exhibit similar 2D structure to GO. Therefore, these new materials
may exhibit similar shape effects and mechanical destruction activ-
ities to GO.
6.2. Exploration of GO-based nanocomposites to achieve synergetic
antibacterial effects
GO exhibits two features in bacterial killing: (1) amphiphilic 2D
structure allowing strong interactions between GO and bacterial
membrane; (2) extracellular MIE-mediated killing mechanisms.
Due to the unique bacterial killing mechanisms that do not involve
intercellular process, GO may have synergetic bactericidal effects
with many conventional antibacterial agents, such as antibiotics,
active biomolecules, silver and inorganic nanoparticles. In addition,
since nitrogen, sulphur, boron or phosphorus atoms on GO may
have significant effect on its surface chemistry that is critical for
bacterial killing, element doping on GO is another promising
method to achieve synergetic bactericidal effects.
6.3. Immobilized GO for antibacterial use
Although GO exhibits significant antibacterial effects, it induces
severe hazard effects in mammals. The toxicity mechanisms in
mammals are very similar to those in bacteria, especially for mem-
brane disruption. Considering the biosafety in real clinical use of
GO, subcutaneous, intravenous or intraperitoneal injections may
be not the appropriate routes of administration. Immobilized GO
as antibacterial nanoproducts for external or non-contact applica-
tions should be encouraged, e.g., additives in filter membrane,
cloth, package materials or coating layer in medical devices.
Conflict of interest
The authors declare that they have no conflict of interest.
Acknowledgments
This work was supported by the National Natural Science Foun-
dation of China (31671032), Key Project of Natural Science Founda-
tion of the Higher Education Institutions of Jiangsu Province
(17KJA310003), and the Priority Academic Program Development
of Jiangsu Higher Education Institutions (PAPD). Ruibin Li was sup-
ported by the Recruitment Program of Global Youth Experts of
China and Strategic Project for Developing Outstanding Institutes
in Suzhou (MCMX201604).
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